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PMID: 116196 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Separation of satellite DNA chromatin and main band DNA chromatin from mouse brain.

Nucleic acids research ·Vol. 7 ·No. 4 ·1979-10-25 ·Pages 935-46

Mazrimas JA, Balhorn R, Hatch FT

Abstract

Using restriction endonucleases which preferentially digest mouse main band DNA and leave satellite DNA intact, we have isolated highly purified chromatin fractions containing only mouse satellite or main band DNA. Following the digestion of mouse brain nuclei with EndoR Alu I, main band DNA chromatin is selectively extracted with 10mM Tris, 10mM EDTA. Satellite DNA chromatin is subsequently extracted from the nuclear pellet with Tris-3M urea and further purified on sucrose gradients. Chromatin extracted from digested nuclei with Tris-EDTA contains only main band DNA and has a molecular weight lower than 2 x 10(6). Chromatin fractions obtained from the lower regions of sucrose gradients of the Tris-Urea extracts contain 40--95% satellite DNA and have a molecular weight of 6 to 8 x 10(6). Both the satellite DNA and main band DNA chromatins contain all five histones and have a protein to DNA ratio of 1.3 to 1.

MeSH Terms
Animals Brain Chemistry Chromatin/isolation & purification Cross-Linking Reagents DNA, Satellite/isolation & purification Formaldehyde Glutaral Histones/isolation & purification Liver/analysis Mice Mice, Inbred Strains
Chemicals
Chromatin Cross-Linking Reagents DNA, Satellite Histones Formaldehyde Glutaral
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Mazrimas J A
Balhorn R
Hatch F T
References (17)
17 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/7.4.935
Published
1979-10-25
Pages
935-46
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342272
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/7.4.935 PMC Full Text View journal details
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